The chelating reaction between glutathione peptides and divalent cadmium ion was used as a typical model for investigating the coordination chemistry of SH-containing peptides and heavy metal ions, which was essential to understand the mechanism of intracellular cadmium detoxification.Here, the mutant (CM113R)7 αHL protein nanopore equipped with a first synthesized per-6-quaternary ammonium-β-cyclodextrin (p-QABCD) was used as nanoreactor and detector to investigate the single-molecule reaction of GSH molecules and Cd2+ ions.Different reaction pathways, intermediates, and products could be recognized.Cd2+-GSH reaction was highly dependent on the solution pH.The Cd(GSH)2 was formed at pH 7.4, while the Cd(GSH)2 and Cd2(GSH)2 were formed at pH 9.0.Cd2(GSH)2 was formed by two possible pathways: (1) A Cd2+ ion primarily coordinated with the thiol group of two GSH molecules to form Cd(GSH)2, and then the second Cd2+ ion quickly incorporated with the deprotonated amino group of Cd(GSH)2 to form Cd2(GSH)2;(2) Two Cd2+ ions separately coordinated with the thiol and deprotonated amino group of one GSH molecule to form Cd2(GSH)1, and the second GSH molecule quickly bounded to Cd2+ ions to form Cd2(GSH)2.This work studied the chelating reaction between metal ions and bioactive glutathione at single-molecule level without labeling and chemical modification, and would be important to further understand intracellular mechanisms of detoxification of heavy metals and greatly expand the research field of nanopore single-molecule technique.

ObjectiveTo evaluate the reliability and validity of the of Disability Assessment Schedule Ⅱ of World Health Organization(WHO-DASⅡ) applied in assessment of the function of disabled athletes.Methods56 disabled people,26 wheelchair basketball athletes and 30 patients,were sampled and completed WHO-DASⅡ questionnaire.The test and re-test had been administrated in 10 days.Barthel Index(BI) and Sports Classification Test had been implemented for the functioning assessment.ResultsThe test-retest reliability for WHO-DASⅡ in six dimensions were: 0.978,0.807,0.938,0.877,0.998,0.935,0.986 and all were in very significant level(P<0.01).There were negative correlation between WHO-DASⅡ scores and BI in total and in every dimension,the correlation coefficients were:-0.77,-0.17,-0.71,-0.82,-0.33,-0.73,-0.61.The correlation coefficients between WHO-DASⅡ total and six dimensional scores and the grade of sports classification were:-0.741,-0.378,-0.806,(-0.541),-0.293,-0.510,-0.677.ConclusionWHO-DASⅡ can be used in assessing the function of disabled athletes.

Objective To investigate the feasibility of transfecting gene into HepG2 by therapeutic ultrasound combined with microbubble,and to explore the optimal ultrasonic parameters for high transfection efficiency.Methods HepG2 cells were cross-interval planted in 24-well plates.EGFP plasmid DNA and microbubbles were added to the cultured HepG2 and three parameters of the therapeutic ultrasound were optimized.Firstly,set ultrasonic intensity gradient (group A,from A1 to A5),which were 0.4 W/cm,0.8 W/cm2,1.2 W/cm2,1.6 W/cm2 and 2.0 W/cm2 respectively.Secondly,set the duty cycle gradient group (group B,from B1 to B3),which were 10％,20％ and 50％ respectively.Lastly,set the exposure time group(group C,from C1 to C3),which were 30 s,90 s and 180 s.After 24 hours,the expression of GFP was observed by fluorescence microscopy,the transfection rates was detected by flow cytometry,the cell death were assessed by trypan blue exclusion.Results The plasmid transfection efficiency was varied under different ultrasonic parameters.Within a certain range,the increasing of parameters can improve the transfection efficiency,but when the parameters were too large (eg.the intensity＞1.6 W/cm2 or 50％ duty cycle),the actual transfection efficiency decreased due to the increased cell death.When the ultrasonic intensity was 1.2 W/cm2,the transfection efficiency and mortality were better than the other subgroups of group A.When the duty cycle was 20％,the transfection efficiency and mortality rate were better than the other subgroups of group B.Continue to increase the exposure time over 90 s was not statistically significant (P ＞0.05) for transfection efficiency and cell mortality.Conclusions The ultrasound-targeted microbubble destruction is an efficient method for gene delivery in vitro.The transfection efficiency vary greatly under different parameters,so optimization parameters is conducive to the promotion of gene transfection.

Objective To investigate the effects of transplantated tumor capillary permeability under microbubble enhanced diagnostic ultrasound exposure.Methods Fourty eight rats were randomly divided into four groups after Walker 256 carcinomas were subcutaneousely implanted,named the control group (A),microbubbles group (B),ultrasound group (C),microbubbles and ultrasound group (D).Using Evans blue(EB) as the indicator for tumor capillary permeability,the microbubbles were injected through the vein with diagnostic ultrasound frequency as 1.75 MHz,mechanical index as 1.4.The spillover of EB was estimated under confocal laser microscope,meanwhile the contents of EB in the tumor tissues were calculated according to the standard curve and spectrophotometry.Results EB spillover was observed around the capillaries in the group D,while EB spillover was only observable on capillary in group A,B and C.The EB content was also significantly higher than those in group A,B and C.Conclusions Exposure to both diagnostic ultrasound and microbubbles results in increased capillary permeability of rat tumor tissues,which might become a potential therapeutic method on clinical chemotherapy.

Objective To compare bolus infusion and replenishment using real low mechanical index contrast enhanced ultrasound in assessing the change of renal cortical perfusion.Methods Using dopamine (i.v.) at the dose of 0.5,2.0,5.0μg · kg- 1 · min- 1 to change renal blood perfusion of 20 rabbits,then during bolus or contant injection of SonoVue,at coded pulse inversion mode,real-time contrast ultrasound was performed,the latter method needed destroying microbubble at a high MI when amplitude reach a steady state,then recording the replenishment,peak intensity(A) and time to peak (PPT) were obtained through raw time-intensity curve,and slope rate of TIC(k) was acquired by curve fitting,standard effective renal plasma flow(ERPF) was measured through 4-aminohippuric acid clearancerate method,meanwhile correlations between ERPF and parameters were analyzed,as well as the paired samples t test for each parameter before and after dopamine administration.Results The ascending branchs of raw TIC of bolus infusion increased sharply and were approximately straight,then descended gradually,while that of replenishment looked like two straightlines with different slopes,then stayed horizontal Both the value of A of two methods were positively correlated with ERPF ( r b =0.85,r re =0.66),and were different at the same ERPF,meanwhile the value of TTP were negatively correlated with ERPF( r b =-0.92,r re =- 0.76),and there were no statistically difference between the two methods.k from Gamma fitting was far from correct,while k from exponential fitting was apparently correlated with ERPF ( r re =0.77 ).Conclusions Both bolus injection and constant injection-replenishment method can assess renal cortical blood perfusion,TIC parameters A and TTP represent regional blood volume fraction and microbubble velocity respectively.Bolus-infusion with real low mechanical index is more precise and available.Comparing with k,TTP is more appropriate to reflect perfusion velocity.

Brain Concussion ; complications ; Hearing ; Hearing Disorders ; prevention & control ; Humans ; Thioctic Acid ; therapeutic use

Hela is the cell line of adenocarcinoma of the uterine cervix, and human papillomavirus (HPV) 18 shows positive. We delivered siRNA with target specifically to HPV18 E7 mRNA into nude mice Hela tumor xenografts by nanopatch to inhibit the HPV gene expression, and further to study the superiority, the best action time and concentration of siRNA of using nanopatch to transfer siRNA in vivo. We designed siRNA that target specifically to HPV18 E7 mRNA (siE7) and checked the effect of siE7 in vitro. Tumor xenografts were transfected with siE7 and GenEscort III by nanopatch. Expression of HPV18 E7 mRNA and protein were detected 0 hours, 24 hours, 48 hours, 72 hours after transfection with PT-PCR and Western blot, and the best action time was analyzed using nanopatch to thansfect siRNA in vivo. We transfected GenEscort III and siE7 of Different concentration into tumor xenografts respectively by nanopatch and intraperitoneal injection. Expression of HPV18 E7 mRNA and protein was detected 72 hours after transfection by PT-PCR and Western blot, to analyze the best action concentration of siRNA and the superiority of using nanopatch to thansfect siRNA in vivo. The results proved that SiE7was efficient to inhibit expression of HPV18 E7 mRNA and to advance Hela apoptosis in vitro. SiE7 transfected by nanopatch into xenografts could inhibit effectively expression of HPV18 E7 mRNA and protein. The best action time and concentration of siRNA of using nanopatch to thansfect siRNA in vivo are 72 hour post-transfection and 2 micromol/L siE7. To compare intraperitoneal injection in delivering siRNA in vivo, the effect of nanopatch is very predominant. It can be well concluded that Nanopatch can effectively transfer siRNA in vivo, which can effectively inhibit the HPV gene expression.
Animals ; Apoptosis ; DNA-Binding Proteins ; genetics ; Female ; Gene Expression Regulation, Viral ; HeLa Cells ; Humans ; Mice ; Mice, Nude ; Nanostructures ; administration & dosage ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; RNA, Small Interfering ; administration & dosage ; Transfection ; Uterine Cervical Neoplasms ; therapy ; Xenograft Model Antitumor Assays

Blue laser imaging (BLI) is a new endoscopic system equipped with the laser beam emitting two different wavelengths.It produces bright and high resolution images for observation of microvascular and microsurface patterns of esophageal and gastric mucosa, helping the diagnosis of early upper gastrointestinal cancer.Compared with the existed endoscopic techniques, BLI shows its unique advantages.In this article, advances in application of BLI in diagnosis of early upper gastrointestinal cancer were reviewed.

AIM:To establish a HPLC fingerprint of Stellera chamaejasme L. from Inner Mongolia Region. METHODS: The RP-HPLC method was used with Akzonobel Kromasil C_ 18 (250 mm?4.6 mm, 5 ?m) the acetonitrile-0.5% phosphoric acid (gradient elution) was used as mobile phase, analytic time was 60 min, and detective wavelength was at 297 nm, the column temperature of 15℃ were adopted. RESULTS: The HPLC fingerprint of Stellera chamaejasme L. set up showed that 14 peaks were co-possessing in different sources. The results of method validation met technical standard of fingerprint, the similarities of Stellera chamaejasme L. were 0.9 to 1.0. CONCLUSION: The method is stable and reliable with a good reproducibility and provides a reference standard for the quality control of Stellera chamaejasme L. from Inner Mongolia Region.

Objective To investigate the feasibility and plan quality of the image-guided volumetric modulated arc therapy (VMAT) based voluntary deep exhale breath-holding technique in the stereotactic ablative body radiotherapy (SABR) for liver tumors.Methods Fifteen patients with liver tumors were involved in this study.All patients were immobilized with voluntary deep exhale breath hold (vDEBH) combined with real-time position management (RPM) respiratory gating system.Treatment was planned using VMAT with 2 modified partial arc and re-planned using intensity modulated radiation therapy (IMRT) technique for comparison.Dosimetric parameters were calculated for plan quality assessment.Quality assurance studies included absolute dose and multiple planar dose verifications,total monitor units and delivery time analysis.Daily cone beam computed tomography imaging was used to verify the motions.Results There were no significant dosimetric differences between VMAT and conventional IMRT plans (P ＞0.05).Both techniques were able to minimize doses to organs at risk including normal liver,kidneys,spinal cord,and stomach.However,the average monitor units with VMAT were significantly lower 28.1％ than those with IMRT(t =3.064,P ＜0.05).The average beam-on time in VMAT plans was 31.6％ shorter than that in IMRT plans(t =2.278,P ＜ 0.05).Conclusions The utilization of VMAT in the treatment planning of SABR for liver tumors under breath control mode has better dosimetrics.In comparison to conventional IMRT plans,VMAT plans have higher efficiency and feasibility.